Titre :
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Phenotypic separation of satellite stem cells using flow cytometry (poster)
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contenu dans :
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Auteurs :
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4th International Congress of Myology, 4ème colloque international de Myologie (9-13 mai 2011; Lille (France)) ;
Briggs D ;
Morgan J ;
Boldrin L
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Type de document :
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Article
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Année de publication :
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2011
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Pages :
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p. 142
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Langues:
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Anglais
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Mots-clés :
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colloque
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Résumé :
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Satellite cells are the principal muscle stem cell, however not all satellite cells contribute equally to muscle regeneration. It is thought that there is a sub-population of satellite cells which is more 'stem cell like' than others. We aim to separate mouse satellite cells into sub-populations and then compare if there is any difference in their ability to proliferate and differentiate into myotubes in vitro and regenerate muscle and self-renew in vivo. Using flow cytometric cell sorting we have separated either freshly isolated or cultured satellite cells on the basis of proliferative state and the level of expression of reactive oxygen species (ROS). Results thus far indicate that using Hoechst and Pyronin Y to separate quiescent satellite cells in G0 phase from those proliferating in G1 is useful for analysis but cells cannot be cultured after sorting due to dye toxicity. Freshly isolated satellite cells have low levels of ROS. Once they are expanded in culture ROS levels increase. Expanded cells were sorted into ROS low, mid and high. ROS low cells have a higher proliferative capacity in vitro than either ROS mid or high. Ongoing experiments will assess whether this translates into the ability to regenerate more muscle in vivo.
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